Flower extract of Panax notoginseng attenuates lipopolysaccharide-induced inflammatory response via blocking of NF-kappaB signaling pathway in murine macrophages.
ABSTRACT The root of Panax notoginseng (PN) is commonly used to treat chronic liver disease with its therapeutic abilities to stop haemorrhage in the circulation, while the PN flower (PN-F) is largely unknown in the biological activities on inflammation and mechanisms of its actions. In this study, the pharmacologic effects of PN-F methanol extract on inflammation were investigated to address potential therapeutic or toxic effects in LPS-stimulated mouse macrophage cells, RAW264.7 cells.
Production of NO, PGE2 and pro-inflammatory cytokines (TNF-alpha and IL-1beta) in supernatant, the expression of iNOS, COX-2 and cytokines, the phosphorylation of MAPK molecules (ERK1/2, JNK and p38 MAPK), and the activation of NF-kappaB in PN-F extract were assayed in LPS-stimulated RAW264.7 cells.
PN-F extract significantly inhibited the productions of NO, PGE2, TNF-alpha and IL-1beta on the LPS-stimulated RAW264.7 cells. In addition, PN-F extract suppressed the mRNA and protein expressions of iNOS, COX-2, TNF-alpha and IL-1beta in LPS-stimulated RAW264.7 cells. The molecular mechanism of PN-F extract-mediated attenuation in RAW264.7 cells has close a relationship to suppressing the phosphorylation of MAPK molecules such as ERK1/2, JNK and p38 MAPK, and the translocation of NF-kappaB p65 subunit into nuclear.
These results indicate that PN-F extract inhibits LPS-induced inflammatory response via the blocking of NF-kappaB signaling pathway in macrophages, and demonstrated that PN-F extract possesses anti-inflammatory properties in vitro.
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ABSTRACT: Allium cepa L. is known to possess numerous pharmacological properties. Our aim was to examine the in vitro effects of Allium cepa L. extract (AcE) on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated by Pg LPS (1μg/mL) and E. coli LPS (1μg/mL) in the presence or absence of different concentrations of AcE (10-1000 μg/mL) for 5 days at 37oC/5% CO2. Resazurin reduction and total protein content assays were used to detect cell viability. AcE did not affect cell viability. Resazurin reduction assay showed that AcE, at up to 1000 μg/mL, did not significantly affect cell viability and cellular protein levels. Additionally a caspase 3/7 luminescence assay was used to disclose apoptosis and there was no difference in apoptotic activity between tested groups and control group. Fluorescence images stained by DAPI showed no alteration on the morphology and cell counts of LPS-stimulated osteoclast precursor cells with the use of AcE in all tested concentrations when compared to control. These findings suggest that Allium cepa L. extract could be used for in vitro studies on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells.International Journal of Cell Biology 06/2014;
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ABSTRACT: Endotoxins from infectious organisms lead to sepsis, a systemic inflammatory response, and a major cause of death. Numerous studies have shown the potential role of plants and plant-derived compounds in the suppression of LPS induced endotoxemia in vivo. In the present study, we have identified a plant namely Seabuckthorn (Hippophae rhamnoides L.) as a potent agent for the treatment of endotoxemia. The objective of the study was to investigate the influence of Supercritical Extract of Seabuckthorn Leaves (SCE200ET) and its active component Isorhamnetin (IR) on the LPS induced endotoxemia in Balb/c mice by measuring the level of nitric oxide (NO), TNF-α and IL-6. Expression of COX-2 and iNOS was measured to understand the involvement of various pathways in the mechanism of action of SCE200ET and IR. The results indicated that SCE200ET and IR inhibited LPS induced NO production by peritoneal macrophages. Cytokines mediated effector functions were influenced by the reduction of IL-6 and TNF-α production and CD40 expression was also markedly diminished in the extract or IR treated groups. In addition, the anti-inflammatory properties were further characterized by decreased expression of COX-2 and iNOS proteins. Fractionation and phytochemical analysis of the extract by RP-HPLC led to identification of isorhamnetin, as bioactive component. Thus, SCE200ET extract and its active component Isorhamnetin could be potential therapeutic agents for the treatment of endotoxin induced sepsis.International immunopharmacology 01/2014; · 2.21 Impact Factor
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ABSTRACT: This study aims to investigate the anti-inflammatory responses and mechanisms of Siegesbeckia orientalis ethanol extract (SOE). In cell culture experiments, RAW264.7 cells were pretreated with SOE and stimulated with lipopolysaccharide (LPS) for inflammatory mediators assay. In animal experiments, mice were tube-fed with SOE for 1 week, and s.c. injected with λ-carrageenan or i.p. injected with LPS to simulate inflammation. The degree of paw edema was assessed, and cytokine profile in sera and mouse survival were recorded. Data showed that SOE significantly reduced NO, IL-6, and TNF-α production in LPS-stimulated RAW264.7 cells. In vivo studies demonstrated that mice supplemented with 32 mg SOE/kg BW/day significantly lowered sera IL-6 level and resulted a higher survival rate compared to the control group (P = 0.019). Furthermore, SOE inhibited LPS-induced NF-κB activation by blocking the degradation of IκB-α. The SOE also reduced significantly the phosphorylation of ERK1/2, p38, and JNK in a dose-dependent manner. In summary, the in vitro and in vivo evidence indicate that SOE can attenuate acute inflammation by inhibiting inflammatory mediators via suppression of MAPKs- and NF-κB-dependent pathways.BioMed Research International 01/2014; 2014:329712. · 2.71 Impact Factor